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Flow cytometric analysis of CD137 (4-1BB) expression on viable stimulated Human peripheral blood lymphocytes. Concanavalin A (Con-A)-stimulated (3 days) peripheral blood mononuclear cells (PBMC) were stained with either BD Horizon™ BUV661 Mouse IgG1, κ Isotype Control (Cat. No. 612966; dashed line histogram) or BD Horizon™ BUV661 Mouse Anti-Human CD137 (4-1BB) antibody (Cat. No. 569682; solid line histogram) at 0.25 µg/test. DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. The fluorescence histogram showing the expression of CD137 (4-1BB) [or Ig Isotype control staining] was derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) activated lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
BD Horizon™ BUV661 Mouse Anti-Human CD137 (4-1BB)
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Preparation and Storage
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BD® CompBeads can be used as surrogates to assess fluorescence spillover (compensation). When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.
For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime BD Horizon Brilliant dyes are used in a multicolor flow cytometry panel. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. When BD Horizon Brilliant Stain Buffer is used in in the multicolor panel, it should also be used in the corresponding compensation controls for all dyes to achieve the most accurate compensation. For the most accurate compensation, compensation controls created with either cells or beads should be exposed to BD Horizon Brilliant Stain Buffer for the same length of time as the corresponding multicolor panel. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).
Note: When using high concentrations of antibody, background binding of this dye to erythroid cell subsets (mature erythrocytes and precursors) has been observed. For researchers studying these cell populations, or in cases where light scatter gating does not adequately exclude these Cells from the analysis, this background may be an important factor to consider when selecting reagents for panel(s).
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関連製品
The 4B4-1 monoclonal antibody specifically binds to CD137 which is also known as 4-1BB, and ILA (induced by lymphocyte activation). CD137 is a type I transmembrane glycoprotein that belongs to the TNF/NGF receptor family. It is encoded by TNFRSF9 (tumor necrosis factor receptor superfamily, member 9). CD137 is expressed on activated T cells, B cells, monocytes, and follicular dendritic cells. CD137 plays roles in the costimulation, differentiation and survival of T cells and B cells.
Development References (6)
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Garni-Wagner BA, Lee ZH, Kim YJ, Wilde C, Kang CY, Kwon BS. 4-1BB is expressed on CD45RAhiROhi transitional T cell in humans. Cell Immunol. 1996; 169(1):91-98. (Immunogen: ELISA, Flow cytometry, Immunoprecipitation). View Reference
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Goodwin, RG. CDw137 (4-1BB) Workshop Panel report. In: Kishimoto T. Tadamitsu Kishimoto .. et al., ed. Leucocyte typing VI : white cell differentiation antigens : proceedings of the sixth international workshop and conference held in Kobe, Japan, 10-14 November 1996. New York: Garland Pub.; 1997:883-884.
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Hurtado JC, Kim SH, Pollok KE, Lee ZH, Kwon BS. Potential role of 4-1BB in T cell activation. Comparison with the costimulatory molecule CD28. J Immunol. 1995; 155(7):3360-3367. (Biology). View Reference
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Kim YJ, Broxmeyer HE. Therapeutic potential of 4-1BB (CD137) as a regulator for effector CD8(+) T cells. J Hematother Stem Cell Res. 2001; 10(4):441-449. (Biology). View Reference
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Zacharakis N, Chinnasamy H, Black M, et al. Immune recognition of somatic mutations leading to complete durable regression in metastatic breast cancer.. Nat Med. 2018; 24(6):724-730. (Clone-specific: Flow cytometry). View Reference
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Zhou Z, Kim S, Hurtado J, et al. Characterization of human homologue of 4-1BB and its ligand. Immunol Lett. 1995; 45(1-2):67-73. (Biology). View Reference
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